Early Stage Alpha-Synuclein Amyloid Fibrils are Reservoirs of Membrane-Binding Species

Abstract The presence of αSN fibrils indisputably associates with the development of synucleinopathies. However, while certain fibril morphologies have been linked to downstream pathological phenotypes, others appear less harmful, leading to the concept of fibril strains, originally described in relation to prion disease. Indeed, the presence of fibrils does not associate directly with neurotoxicity. Rather, it has been suggested that the toxic compounds are soluble amyloidogenic oligomers, potentially co-existing with fibrils. Here, combining synchrotron radiation circular dichroism, transmission electron microscopy and binding assays on native plasma membrane sheets, we reveal distinct biological and biophysical differences between initial and matured fibrils, transformed within the timespan of few days. Immature fibrils are reservoirs of membrane-binding species, which in response to even gentle experimental changes release into solution in a reversible manner. In contrast, mature fibrils, albeit macroscopically indistinguishable from their less mature counterparts, are structurally robust, shielding the solution from the membrane active soluble species. We thus show that particular biological activity resides transiently with the fibrillating sample, distinct for one, but not the other, spontaneously formed fibril polymorph. These results shed new light on the principles of fibril polymorphism with consequent impact on future design of assays and therapeutic development

Photoionization of High Altitude Gas in a Supernova-Driven Turbulent Interstellar Medium

We investigate models for the photoionization of the widespread diffuse ionized gas in galaxies. In particular we address the long standing question of the penetration of Lyman continuum photons from sources close to the galactic midplane to large heights in the galactic halo. We find that recent hydrodynamical simulations of a supernova-driven interstellar medium have low density paths and voids that allow for ionizing photons from midplane OB stars to reach and ionize gas many kiloparsecs above the midplane. We find ionizing fluxes throughout our simulation grids are larger than predicted by one dimensional slab models, thus allowing for photoionization by O stars of low altitude neutral clouds in the Galaxy that are also detected in Halpha. In previous studies of such clouds the photoionization scenario had been rejected and the Halpha had been attributed to enhanced cosmic ray ionization or scattered light from midplane H II regions. We do find that the emission measure distributions in our simulations are wider than those derived from Halpha observations in the Milky Way. In addition, the horizontally averaged height dependence of the gas density in the hydrodynamical models is lower than inferred in the Galaxy. These discrepancies are likely due to the absence of magnetic fields in the hydrodynamic simulations and we discuss how magnetohydrodynamic effects may reconcile models and observations. Nevertheless, we anticipate that the inclusion of magnetic fields in the dynamical simulations will not alter our primary finding that midplane OB stars are capable of producing high altitude diffuse ionized gas in a realistic three-dimensional interstellar medium.Comment: ApJ accepted. 17 pages, 7 figure

Spatiotemporal Dynamics of Mediterranean Shallow Coastal Fish Communities Along a Gradient of Marine Protection

The importance of habitat factors in designing marine reserves and evaluating their performance over time has been regularly documented. Over three biennial sampling periods, we examined the effects of vegetated coverage and habitat diversity (i.e., patchiness) on fish density, community composition, and species-specific patterns along a gradient of protection from harvest in the shallow Spanish southern Mediterranean, including portions of the Tabarca marine reserve. With the exception of two herbivores (Sarpa salpa and Symphodus tinca), vegetated cover did not significantly affect fish densities, while habitat diversity was an influential factor across all three sampling periods. Overall, fish density was more positively associated with more continuous vegetated or unvegetated habitats, and was greatest in areas of highest protection (Tabarca II – Isla Nao site). These patterns were usually observed for four abundant fish species (Boops boops, Chromis chromis, Oblada melanura, and S. salpa). Fish community composition was distinct in the most protected portion of the Tabarca reserve, where it was also most stable. Our findings align with previous investigations of the Tabarca reserve and its surrounding areas, and demonstrate its continued effectiveness in conserving fish biomass and habitat. Together with effective management, marine reserves can facilitate greater species abundance, more stable biological communities, and resilient ecosystems

Estimating the Porosity of the Interstellar Medium from Three-Dimensional Photoionization Modeling of H II Regions

We apply our three dimensional photoionization code to model Wisconsin H-alpha Mapper observations of the H II region surrounding the O9.5V star Zeta Oph. Our models investigate the porosity of the interstellar medium around zeta Oph and the effects of 3D densities on the H-alpha surface brightness and variation in the [N II]/H-alpha line ratio. The Zeta Oph H II region has a well characterized ionizing source, so it is an excellent starting point for 3D models of diffuse ionized gas. We investigate various hierarchically clumped density structures, varying the overall smoothness within the clumping algorithm. By simulating the observations, we can estimate the porosity of the medium in the vicinity of Zeta Oph and find that within the context of our hierarchically clumped models, around 50% to 80% of the volume is occupied by clumps surrounded by a low density smooth medium. We also conclude that in order for O stars to ionize the diffuse Warm Ionized Medium, the O star environment must be more porous than that surrounding Zeta Oph, with clumps occupying less than one half of the interstellar volume. Our clumpy models have irregular boundaries, similar to observed H II regions. However, in observed H II regions it is difficult to identify the precise location of the boundary because of the foreground and/or background emission from the widespread Warm Ionized Medium. This complicates the interpretation of the predicted rapid rise of some emission line ratios near the edge of uniform density H II regions and combined with the three dimensional clumpy nature of the interstellar medium may explain the apparent lack of distinctive emission line ratios near H I -- H II interfaces.Comment: Accepted for publication in Ap

Interaction between the glutamate transporter GLT1b and the synaptic PDZ domain protein PICK1

This is the published version. Copyright WileySynaptic plasticity is implemented by the interaction of glutamate receptors with PDZ domain proteins. Glutamate transporters provide the only known mechanism of clearance of glutamate from excitatory synapses, and GLT1 is the major glutamate transporter. We show here that GLT1 interacts with the PDZ domain protein PICK1, which plays a critical role in regulating the expression of glutamate receptors at excitatory synapses. A yeast two-hybrid screen of a neuronal library using the carboxyl tail of GLT1b yielded clones expressing PICK1. The GLT1b C-terminal peptide bound to PICK1 with high affinity (Ki = 6.5 ± 0.4 μm) in an in vitro fluorescence polarization assay. We also tested peptides based on other variants of GLT1 and other glutamate transporters. GLT1b co-immunoprecipitated with PICK1 from rat brain lysates and COS7 cell lysates derived from cells transfected with plasmids expressing PICK1 and GLT1b. In addition, expression of GLT1b in COS7 cells changed the distribution of PICK1, bringing it to the surface. GLT1b and PICK1 co-localized with each other and with synaptic markers in hippocampal neurons in culture. Phorbol ester, an activator of protein kinase C (PKC), a known PICK1 interactor, had no effect on glutamate transport in rat forebrain neurons in culture. However, we found that exposure of neurons to a myristolated decoy peptide with sequence identical to the C-terminal sequence of GLT1b designed to block the PICK1–GLT1b interaction rendered glutamate transport into neurons responsive to phorbol ester. These results suggest that the PICK1–GLT1b interaction regulates the modulation of GLT1 function by PKC.The authors are grateful to Sara Vasquez who provided excellent technical assistance in preparing the neuronal cultures. In addition, we are grateful for helpful discussions with Drs Gabriel Corfas, Michael Berne and Michael Robinson, to Dr Tom Schwarz for reading an early version of this manuscript, and to Dr Jeff Rothstein for providing an anti-cGLT1a antibody. We are also indebted to Dr Robinson for providing us with a detailed protocol for the biotinylation studies. This work was funded by grants from the Ron Shapiro Charitable Foundation (P.A.R.), the Muscular Dystrophy Association (P.A.R.), and National Institutes of Health research grant NS 40753 and a Mental Retardation Core Grant HD18655

A multicenter, community-based, mixed methods assessment of the acceptability of a triple drug regimen for elimination of lymphatic filariasis

BackgroundMany countries will not reach elimination targets for lymphatic filariasis in 2020 using the two-drug treatment regimen (diethylcarbamazine citrate [DEC] and albendazole [DA]). A cluster-randomized, community-based safety study performed in Fiji, Haiti, India, Indonesia and Papua New Guinea tested the safety and efficacy of a new regimen of ivermectin, DEC and albendazole (IDA).Methodology/principal findingsTo assess acceptability of IDA and DA, a mixed methods study was embedded within this community-based safety study. The study objective was to assess the acceptability of IDA versus DA. Community surveys were performed in each country with randomly selected participants (>14 years) from the safety study participant list in both DA and IDA arms. In depth interviews (IDI) and focus group discussions (FGD) assessed acceptability-related themes. In 1919 individuals, distribution of sex, microfilariae (Mf) presence and circulating filarial antigenemia (CFA), adverse events (AE) and age were similar across arms. A composite acceptability score summed the values from nine indicators (range 9-36). The median (22.5) score indicated threshold of acceptability. There was no difference in scores for IDA and DA regimens. Mean acceptability scores across both treatment arms were: Fiji 33.7 (95% CI: 33.1-34.3); Papua New Guinea 32.9 (95% CI: 31.9-33.8); Indonesia 30.6 (95% CI: 29.8-31.3); Haiti 28.6 (95% CI: 27.8-29.4); India 26.8 (95% CI: 25.6-28) (PConclusions/significanceIDA and DA regimens for LF elimination were highly and equally acceptable in individuals participating in the community-based safety study in Fiji, Haiti, India, Indonesia, and Papua New Guinea. Country variation in acceptability was significant. Acceptability of the professionalism of the treatment delivery was highlighted